title: A Penicillium sp. F33 metabolite and its synthetic derivatives inhibit acetyl-CoA:1-O-alkyl-sn-glycero-3-phosphocholine acetyltransferase (a key enzyme in platelet-activating factor biosynthesis) and carrageenan-induced paw edema in mice.
abstract: Acetyl-CoA:1-O-alkyl-sn-glycero-3-phosphocholine (lyso-PAF) acetyltransferase is a key enzyme in the biosynthesis of 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine (PAF) in inflammatory cells. Substances which inhibit this enzyme are of therapeutic interest. In this study, we screened for new inhibitors of lyso-PAF acetyltransferase with anti-inflammatory effects. In a metabolite from Penicillium sp. F33, we isolated an acetyltransferase inhibitor identified as dihydrofumigatin (2-methoxy-1,3,4-trihydroxy-5-methylbenzene) from high resolution mass spectrometer and NMR data. Dihydrofumigatin had strong acetyltransferase inhibitory activity, but was not stable in aqueous solution. Thus, we chemically synthesized its oxidized form fumigatin (3-hydroxy-2-methoxy-5-methyl-1,4-benzoquinone) and derivatives thereof, and evaluated their inhibitory effects. Strong inhibitory activity was observed for saturated fatty acid esters of fumigatin; the order of inhibition was 3-decanoyloxy-2-methoxy-5-methyl-1,4-benzoquinone (termed FUD-7, IC₅₀ = 3 μM)>2-methoxy-5-methyl-3-tetradecanoyloxy-1,4-benzoquinone (termed FUD-8, IC₅₀ = 20 μM)>3-hexanoyloxy-2-methoxy-5-methyl-1,4-benzoquinone (IC₅₀ = 139 μM). Interestingly, these compounds also significantly suppressed the gene expression of lyso-PAF acetyltransferase/LPCAT2 in mouse bone marrow-derived macrophages stimulated by lipopolysaccharide (LPS). We further evaluated the effect of these substances on anti-inflammatory activity in vivo using the carrageenan-induced mouse paw edema test. FUD-7 and FUD-8 at 2.5 mg/kg showed significant, 47.9-51.7%, inhibition stronger than that of prednisolone at 10 mg/kg (41.9%). These results suggest that FUD-7 and FUD-8 are potent inhibitors with anti-inflammatory activity.
authors.affiliation: Department of Pharmaco-Biochemistry, School of Pharmaceutical Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka 422-8526, Japan.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Yamazaki
authors.forename: Kengo
authors.initials: K
authors.lastname: Yasuda
authors.forename: Tensei
authors.initials: T
authors.lastname: Matsuyama
authors.forename: Takuya
authors.initials: T
authors.lastname: Ishihara
authors.forename: Ryoko
authors.initials: R
authors.lastname: Higa
authors.forename: Taira
authors.initials: T
authors.lastname: Sawairi
authors.forename: Masahiko
authors.initials: M
authors.lastname: Yamaguchi
authors.forename: Masahiro
authors.initials: M
authors.lastname: Egi
authors.forename: Shuji
authors.initials: S
authors.lastname: Akai
authors.forename: Toshio
authors.initials: T
authors.lastname: Miyase
authors.forename: Akira
authors.initials: A
authors.lastname: Ikari
authors.forename: Masao
authors.initials: M
authors.lastname: Miwa
authors.forename: Junko
authors.initials: J
authors.lastname: Sugatani
mesh_terms: D000123:Acetyltransferases / Q000037:antagonists & inhibitors*; D000818:Animals; D001483:Base Sequence; D016227:Benzoquinones / Q000494:pharmacology; D002351:Carrageenan / Q000633:toxicity*; D002478:Cells, Cultured; D003512:Cyclohexanones / Q000494:pharmacology; D017931:DNA Primers; D004487:Edema / Q000139:chemically induced / Q000201:enzymology*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D010407:Penicillium / Q000378:metabolism*; D010972:Platelet Activating Factor / Q000096:biosynthesis*; D051381:Rats; D017208:Rats, Wistar; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D016227:Benzoquinones; D003512:Cyclohexanones; D017931:DNA Primers; D010972:Platelet Activating Factor; D002351:Carrageenan; C018067:fumigatin; D000123:Acetyltransferases; C046035:1-alkylglycerophosphocholine acetyltransferase
keywords: 1,4-diacetoxy-2-methoxy-5-methyl-3-tetradecanoyloxybenzene; 1-O-alkyl-sn-glycero-3-phosphocholine; 1-hexadecyl-2-N-methylcarbamyl-sn-glycero-3-phosphocholine; 2-methoxy-5-methyl-1,3,4-triacetoxybenzene; 2-methoxy-5-methyl-3-octadecanoyloxy-1,4-benzoquinone; 2-methoxy-5-methyl-3-tetradecanoyloxy-1,4-benzoquinone; 3,4-dihydroxy-1,2-dimethoxy-5-methylbenzene; 3-acetoxy-2-methoxy-5-methyl-1,4-benzoquinone; 3-decanoyloxy-2-methoxy-5-methyl-1,4-benzoquinone; 3-hexanoyloxy-2-methoxy-5-methyl-1,4-benzoquinone; 3-hydroxy-2-methoxy-5-methyl-1,4-benzoquinone (fumigatin); BMDMs; Carrageenan-induced edema; DART; FUD-1; FUD-2; FUD-3; FUD-4; FUD-5; FUD-6; FUD-7; FUD-8; FUD-9; Fumigatin; HPLC; HRMS; LPCAT2; LPS; Lyso-PAF acetyltransferase; NDGA; PAF; PAF biosynthesis; Platelet activating factor; TCA; TLC; bone marrow-derived macrophages; c-PAF; direct analysis in real time; high resolution mass spectrometer; high-performance liquid chromatography; lipopolysaccharide; lyso-PAF; lysophosphatidylcholine acyltransferase 2; nordihydroguaiaretic acid; platelet-activating factor, 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine; thin layer chromatography; trichloroacetic acid